Regeneration of spermatogenesis by grafting testicular tissue or injecting testicular cells into the testes of sterile mice: a comparative study  Dorien Van Saen, M.Sc., Ellen Goossens, Ph.D., Gert De Block, Herman Tournaye, M.D., Ph.D.  Fertility and Sterility  Volume 91, Issue 5, Pages 2264-2272 (May 2009) DOI: 10.1016/j.fertnstert.2008.02.100 Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Schematic presentation of the determination of the tubule length. (Left) The diameter of one microscopic view at magnification ×100 was measured and found to be 2 mm. (Middle) After SSCT, GFP+ tubules were present next to GFP− tubules. The length of the GFP+ tubules was measured by following the positive tubules and determining how many diameters they were long (length = number of diameters × 2 mm). (Right) After grafting the entire microscopic view was green. On average three tubules were visible in one microscopic view, which means 6 mm of tubules. The amount of microscopic views was counted and multiplied by 6 (length = number of microscopic views × 6 mm). (B) Testis of a 5-day-old GFP+ mouse under UV light. The green fluorescence is only situated on the wall, whereas there is no fluorescence in the lumen. (C,D) GFP+—colonies after ductal injection. (E) GFP+—cells after blind injection. (F) GFP+—colonies after blind injection. (G) GFP+—colony after graft. Fertility and Sterility 2009 91, 2264-2272DOI: (10.1016/j.fertnstert.2008.02.100) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Comparison between weight and colony length of testes after ductal injection, blind injection, and grafting (mean ± SD). Weight: P<.0001 by Mann Whitney (ductal injection − grafting); P=.0077 by Mann Whitney (blind injection − grafting). Colony length: P=.0033 by Mann Whitney (ductal injection − grafting); P=.0003 by Mann Whitney (blind injection − grafting). Fertility and Sterility 2009 91, 2264-2272DOI: (10.1016/j.fertnstert.2008.02.100) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Anti-GFP staining. (A) negative control: GFP− testis. (B) positive control: GFP+ testis. (C) Tubules with donor spermatogenesis after SSCT. (D) Tubules with donor spermatogenesis after graft. (E) Donor-derived spermatogenesis after grafting fresh tissue. (F) Donor-derived spermatogenesis after grafting frozen-thawed tissue. (G) Sclerosis after grafting. Fertility and Sterility 2009 91, 2264-2272DOI: (10.1016/j.fertnstert.2008.02.100) Copyright © 2009 American Society for Reproductive Medicine Terms and Conditions